Conformational Dynamics of the Guanidine-IV Riboswitch Reveal a Ligand-Sensitive Transcriptional Window

Temel Kavramlar

The guanidine-IV riboswitch exhibits a narrow transcriptional window that is highly sensitive to guanidine binding, with the aptamer domain playing a crucial role in ligand sensing and regulating gene expression.

Özet

This study investigated the conformational dynamics of the guanidine-IV riboswitch upon ligand binding using single-molecule fluorescence resonance energy transfer (smFRET). The key findings are:

The aptamer domain of the guanidine-IV riboswitch exhibits higher sensitivity to guanidine compared to the terminator and full-length constructs. Magnesium ions (Mg2+) and guanidine (Gua+) facilitate the formation of a kissing loop (KL) in the aptamer domain, significantly increasing the KL-formed conformation.

In contrast, the addition of Gua+ and Mg2+ only had a minimal effect on the formation of the KL-formed anti-terminator structure in the terminator/anti-terminator or full-length RNA.

By mimicking the transcriptional process using position-specific labeling of RNA (PLOR) and smFRET, the authors discovered a narrow Gua+-sensitive transcriptional window for the guanidine-IV riboswitch, with a dramatic structural switch observed during the extension from 87 to 88 nucleotides upon Gua+ addition.

The authors propose a folding model for the guanidine-IV riboswitch, where the competition between the P2 stem and the KL structure determines the proportions of the terminator and the anti-terminator conformations as transcription progresses.

Overall, the study provides valuable insights into the ligand-response mechanism of the guanidine-IV riboswitch, highlighting the crucial role of the aptamer domain in sensing and regulating gene expression.

İstatistikler

The EC50 value for the transcriptional read-through of the guanidine-IV riboswitch is 68.7 ± 7.3 μM.
The Kd for guanidine binding to the riboG-apt in the presence of 0.5 mM Mg2+ is 286.0 ± 18.1 μM.

Alıntılar

"The aptamer domain of the guanidine-IV riboswitch exhibits a greater sensitivity to guanidine, surpassing that observed in both the terminator and full-length riboswitch."
"A finely-tuned transcriptional window of the guanidine-IV riboswitch was detected to be responsive to ligand binding."
"Furthermore, a folding-function model for the guanidine-IV riboswitch under both guanidine-free and guanidine-present conditions offers valuable insights into its regulatory mechanism."

Önemli Bilgiler Şuradan Elde Edildi

Ligand Response of Guanidine-IV riboswitch at Single-molecule Level

by Gao,L., Chen... : www.biorxiv.org 12-05-2023

https://www.biorxiv.org/content/10.1101/2023.12.05.570130v2

Daha Derin Sorular

What are the potential applications of the guanidine-IV riboswitch in synthetic biology or biotechnology, given its unique ligand-sensing capabilities

The guanidine-IV riboswitch holds significant potential in synthetic biology and biotechnology due to its unique ligand-sensing capabilities. One potential application lies in the development of novel biosensors for detecting guanidine levels in various biological samples. By utilizing the specific binding affinity of the guanidine-IV riboswitch for guanidine, researchers can engineer biosensors that provide real-time, sensitive detection of guanidine concentrations. This could be particularly valuable in environmental monitoring, medical diagnostics, and bioproduction processes where guanidine levels need to be monitored and controlled.
Another application could be in the design of genetic circuits for controlling gene expression in response to guanidine levels. By incorporating the guanidine-IV riboswitch into genetic circuits, researchers can create systems that regulate the expression of target genes based on the presence or absence of guanidine. This could be utilized in the development of smart therapeutics, bioremediation strategies, or metabolic engineering approaches where precise control over gene expression is required.

How do the structural differences between the guanidine-IV riboswitch and other guanidine riboswitches contribute to their distinct regulatory mechanisms

The structural differences between the guanidine-IV riboswitch and other guanidine riboswitches play a crucial role in their distinct regulatory mechanisms. The guanidine-IV riboswitch differs significantly from other guanidine riboswitches in its ability to enhance transcription of specific genes by forming an anti-terminator structure upon guanidine binding. This unique mechanism allows the guanidine-IV riboswitch to regulate gene expression in response to guanidine levels, facilitating the export of guanidine and reducing its toxicity in cells.
In contrast, other guanidine riboswitches may function through different structural motifs or regulatory mechanisms, leading to diverse outcomes in gene expression. For example, the guanidine-I riboswitch may regulate gene expression by forming alternative secondary structures or interacting with different regulatory proteins. These variations in structure and function contribute to the specificity and versatility of guanidine riboswitches in sensing and responding to guanidine in the cellular environment.

Could the insights gained from the guanidine-IV riboswitch be applied to understand the co-transcriptional folding and ligand-sensing mechanisms of other classes of riboswitches

The insights gained from studying the guanidine-IV riboswitch could indeed be applied to understand the co-transcriptional folding and ligand-sensing mechanisms of other classes of riboswitches. The co-transcriptional folding of RNA plays a critical role in its function, especially in riboswitches involved in gene regulation. By investigating the conformational dynamics of the guanidine-IV riboswitch during transcription, researchers can gain valuable insights into how RNA structures form and change in response to ligand binding.
These insights can be extrapolated to other classes of riboswitches to elucidate their co-transcriptional folding pathways, ligand-sensing behaviors, and regulatory mechanisms. By applying similar methodologies and techniques used in studying the guanidine-IV riboswitch, researchers can uncover the structural transitions and ligand-induced changes in various riboswitches, providing a deeper understanding of their functions and potential applications in synthetic biology, biotechnology, and medicine.